Standard samples of volatiles, including octanal (purity≥99%), methyl salicylate (analytical standard), undecane (analytical standard), hexadecane (analytical standard), and tridecanal (purity≥95%) were purchased from the Sigma-Aldrich Corporation. Using liquid paraffin as a solvent, solutions diluted 102, 104 and 106 times were prepared for testing. Internal standards n-octane and nonyl acetate were purchased from Sigma-Aldrich; 400 ng of octane and 400 ng of nonyl acetate were weighed and mixed with 20 µL hexane as the internal standard.
Nonyl acetate
Manufactured by Merck Group
Sourced in United States
Nonyl acetate is a clear, colorless liquid chemical compound. It is used as a laboratory reagent and as a component in various industrial applications.
Automatically generated - may contain errors
Lab products found in correlation
Octanal, by Merck Group (1 mentions)
N octane, by Merck Group (1 mentions)
Hexadecane, by Merck Group (1 mentions)
Methyl salicylate, by Merck Group (1 mentions)
Undecane, by Merck Group (1 mentions)
Tridecanal, by Merck Group (1 mentions)
Qp 2010 gc ms instrument, by Shimadzu (1 mentions)
N hexane, by Tedia (1 mentions)
N hexanal, by Merck Group (1 mentions)
Hp 5ms capillary column, by Agilent Technologies (1 mentions)
Intuvo 9000 gc system, by Agilent Technologies (1 mentions)
C8 c20 hydrocarbon standard, by Merck Group (1 mentions)
Thermomixer, by Eppendorf (1 mentions)
Methyl tert butyl ether (mtbe), by Merck Group (1 mentions)
Hexane, by Merck Group (1 mentions)
10 protocols using nonyl acetate
1
Volatile Compound Analysis Using Diluted Solutions
2
Extraction of Plant Root Metabolites
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For each plant, the entire tap root system was collected in the field and transported to the lab. Fresh root tissue was ground into powder in liquid nitrogen. Ethyl acetate (Macklin Technology, Shanghai, China) was added to the powder in a 5:1 (volume to weight) ratio, with nonyl acetate (CAS:143-13-5, ≥98%, Sigma Aldrich, St Louis, MO, USA) included (0.002%) as an internal standard. After shaking at 200 rpm at room temperature for two hours and centrifugation at 5000 rpm for five minutes, the organic phase was collected for subsequent chemical analysis and bioassays.
3
Analysis of Rice Plant Volatiles
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Plant VOCs were collected with a closed loop dynamic headspace sampling system, similar to the method described by Sun et al.59 (link). One milliliter of n-Hexane (Tedia, USA) was applied to elute VOCs from absorbent traps and 1300 ng Nonyl acetate (Sigma, Switzerland) was added to each sample as an internal standard. VOCs emitted from the Xoo infected rice and control rice plants were collected for 24 h (16 h in light and 8 h in darkness) at room temperature; BPH-induced plant volatiles from rice plants (each rice plant was also infested with 15 gravid female adults for 24 h) were collected for 8 h in the light (20000 lx) as described by Lou et al.60 (link). Each treatment contained three to five biological replicates.
Gas chromatography–mass spectrometry (GC/MS) analyses of VOCs were performed on a QP-2010 GC/MS instrument (Shimadzu, Japan) equipped with an HP-5 MS fused-silica column (30 m × 0.25 mm × 0.25 μm). (Agilent Technologies,http://www.agilent.com ). Helium (1 ml/min) was used as the carrier gas, and the initial oven temperature was 40 °C, held for 1 min, ramped at 8 °C min−1 to 300 °C held for 5 min. VOCs were identified by comparing their GC retention indices and MS spectra with those from the NIST11 library. The Retention index for each compound was determined using a series of straight chain alkanes (C7-C30) as standards.
Gas chromatography–mass spectrometry (GC/MS) analyses of VOCs were performed on a QP-2010 GC/MS instrument (Shimadzu, Japan) equipped with an HP-5 MS fused-silica column (30 m × 0.25 mm × 0.25 μm). (Agilent Technologies,
4
Organic Extraction and GC-MS Analysis
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For organic extraction, leaf tissue of each cultivar was ground into powder and ethyl acetate of HPLC grade (Macklin Technology, Shanghai, China) was added as solvent in 5:1 (volume to weight) ratio. After shaking at room temperature for 2 h and a subsequent centrifugation (5000 r/min, 5 min), the organic phase was collected for GC-MS analysis and antifungal bioassays. For preparation of samples for GC-MS analysis, nonyl acetate (CAS:143-13-5, ≥98%, Sigma Aldrich, St Louis, MO, USA) was added to ethyl acetate (0.002%) as an internal standard. For each cultivar, three biological replicates were analyzed.
5
Extraction of Organic Compounds
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Fresh leaf or flowerhead tissue (0.2 g) was ground into powder in liquid nitrogen. Ethyl acetate (Macklin Technology, Shanghai, China) was added into the powder in a 5:1 (volume to weight) ratio, with nonyl acetate (CAS: 143-13-5, ≥98%, Sigma-Aldrich, Saint Louis, MO, USA) included (0.002%) as an internal standard. After shaking at 200 rpm at room temperature for two hours and centrifugation at 5000 rpm for five minutes, the organic phase was collected for the following chemical analysis and bioassays.
6
Synthesis and Characterization of Aldehydes
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(E)-2-hexen-1-al (E2al), was purchased from Bedoukian (Bedoukian Research, Danbury, CT, USA). (Z)-3-hexen-1-al (Z3al), n-Hexanal (nHal), and nonyl acetate were purchased from Sigma-Aldrich (St. Louis, MO, USA). All solvents used were analytical grade.
7
Profiling Fungal Volatile Metabolites
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Actively growing fungi (Supplementary Table S1 ) were inoculated on PDA, and headspace volatiles were collected after 4 days of incubation in darkness at 25 °C. A single Petri dish (control or fungus inoculated) was placed inside a 1 L volatile collection glass jar with the Petri dish lid open slightly (~5 mm) to facilitate diffusion of volatiles. Active charcoal-filtered air was pumped into the collection jar and the outgoing air was passed through a filter packed with 150 mg Super Q adsorbent at a flow rate of 0.45 L min−1 for 24 h. After each experiment, the surface margin of fungus on the PDA plate was photographed and measured using ImageJ software. Volatile traps were eluted twice with 200 µL dichloromethane spiked with 10 ng µL−1 nonyl acetate (Sigma Aldrich) as an internal standard.
8
GC-MS Analysis of Terpenoid Compounds
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Terpenoid compounds were analyzed using a GC-MS system (Agilent Intuvo 9000 GC system coupled with an Agilent 7000 Triple Quadrupole mass detector), equipped with an Agilent HP 5 MS capillary column (30 nm × 0.25 mm); helium (99.99%) was used as the carrier gas at a flow rate of 1 mL/min. The injection volume of each sample was 1 µL. The temperature of the injection port was 260 °C, with a spilt mode (spilt ratio = 5:1) at the rate of 5 °C/min. The column oven temperature program was as follows: the temperature was initiated at 40 °C, followed by an increase to 250 °C at a rate of 5 °C/min. The MS condition included an EI ion source temperature of 230 °C, ionization energy of 70 eV and mass scan range of 40–500 amu. The separated constituents were identified using the NIST17 MS library (National Institute of Standards and Technology). A C8-C20 hydrocarbon standard (Sigma-Aldrich, St. Louis, MO, USA) was used to obtain the retention indices. Each constituent was quantified based on the comparison of its peak area with that of the internal standard (Nonyl acetate, Sigma-Aldrich, St. Louis, MO, USA), and the contents were expressed as µg g−1 fresh weight.
9
Volatile Extraction from Strawberry Fruit
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Strawberry fruit from transient expression assay were quenched in liquid nitrogen immediately upon harvest and subsequently stored at −80 °C prior to extraction. Frozen berries were crushed and then ground to a fine powder in a liquid nitrogen-cooled coffee grinder (KitchenAid Blade Coffee Grinder, St. Joseph, MI). Volatiles extractions were carried out in technical triplicate for each sample. Volatile compounds were extracted from samples, 250 mg of finely ground powder, using 1 mL of MTBE (Sigma-Aldrich) containing nonyl acetate (Sigma-Aldrich) at 500 ng mL−1 as a surrogate standard. Samples were vortexed for 10 s then shaken for 15 min at 1400 RPM at 25 °C in a Thermomixer (Eppendorf). Following 5 min of centrifugation at 4000 x g the MTBE phase was transferred into a glass sample vial. All samples were evaporated under nitrogen gas to a volume of 100 μL for volatile analysis.
10
Headspace Volatile Collection of Tobacco Plants
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The collection of feeding-induced volatiles by M. persicae on tobacco plants was conducted using the headspace absorption method in a static environment. The whole tobacco plant was deposited in a 30×50 cm 2 polyacetate bag for volatiles collection (Anfora et al., 2009) . The charcoal-filtered air from the headspace of the bag was pushed at a rate of 200 mL/min through a sorbent column (50 mg Porapak Type Q, Sigma-Aldrich) connected to an air pump through Teflon tubing. The volatile collections for each treatment were replicated for three times, according to the same protocol. The collected volatiles were eluted from the sorbent column using 500 μL of hexane (purity >99%, Sigma-Aldrich) as solvent desorption at room temperature. The collections were concentrated to 50 μL by slow nitrogen stream, and 0.5 μg of nonyl acetate (purity ≥99%, Sigma-Aldrich) was added as the internal standard. The extracts were preserved in 2-mL brown vials at -20℃ until utilized for the GC-MS analysis.
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